CN1634925A - Separation and purification method for ginkgolide monomer - Google Patents
Separation and purification method for ginkgolide monomer Download PDFInfo
- Publication number
- CN1634925A CN1634925A CNA2004100651047A CN200410065104A CN1634925A CN 1634925 A CN1634925 A CN 1634925A CN A2004100651047 A CNA2004100651047 A CN A2004100651047A CN 200410065104 A CN200410065104 A CN 200410065104A CN 1634925 A CN1634925 A CN 1634925A
- Authority
- CN
- China
- Prior art keywords
- bilobalide
- column chromatography
- supercritical
- solvent
- ginkgolide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Landscapes
- Medicines Containing Plant Substances (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
A separation and purification process for bilobalide monomers is provided. The column chromatography-recrystallization-supercritical CO2 refinement purification combined process is adopted to prepare high purity bilobalide monomers. The column chromatography adopts positive or reverse fixed packing material. Water and ethyl acetate are used for gradient elution. Supercritical CO2 is used for refinement purification and crystallographic form arrangement. The process conditions are: the pressure is less than or equal to 100Mpa, and the temperature is less than or equal to 150 Celsius degrees. The obtained monomers content amounts to 99.8% at the utmost, and the total yield is greater than or equal to 90%.
Description
One, technical field
The present invention relates to a kind of in plant milk extract the method for the various monomer reactivity compositions of separation and purification, exactly be separation and purification bilobalide-like monomer methods in Folium Ginkgo extract.
Two, background technology
The one-tenth of Ginkgo Leaf is grouped into extremely complicated, includes the various active composition, mainly contains flavones, terpene lactones, also has organic acid, alkylphenol and alkyl phenolic acid, steroidal compounds, trace element etc. in addition.Medicinal ingredients in the Ginkgo Leaf mainly is ginkgolic flavone glycoside class and bilobalide-like substances.
Folium Ginkgo extract (GBE) has been listed Pharmacopoeia of People's Republic of China (2000 version, enlarged edition) in.Press the medicinal GBE of this standard production at present, Ginkgo total flavones content 〉=24%, Ginkgolides content 〉=6%.
Ginkgolides comprises Ginkgolide A, B, C, J, M and bilobalide etc., belongs to the ginkgo endemic element.Be called the bilobalide-like monomer in this application.According to the literature, each monomeric pharmic function has certain difference in the Ginkgolides.Ginkgolide A, B have significant curative effect to the treatment cardiovascular and cerebrovascular diseases.Bilobalide is to treating then drug effect uniqueness of nervous system disorders.For improving result of treatment, control drug quality, the development of new preparation sometimes must be with each monomer separation purifying.
Bilobalide is that sesquiterpene, Ginkgolide A, B, C, J, M are the diterpenes materials.Each monomeric molecular structure is comparatively similar, and polarity difference is very little, the extraction that causes ginkgolide monomer with separate the comparison difficulty.
Be seen in the ginkgolide monomer of bibliographical information and mainly contain alcohols extraction-extraction-column chromatography, alcohols extraction-extraction-reversed phase chromatography separation method, supercritical fluid extraction (SCFE) etc.
Day disclosure special permission communique, flat 3-24084 (1991) has introduced solvent extraction-lead acetate precipitation-silica gel adsorption-recrystallization method.Its bilobalide extraction process flow process is as follows:
Extraction weeding of grease soluble substance, extracting phase is with low alkyl group acetic ester (or lower ketones, benzene) extraction, with organic phase precipitation drying, after the aqueous phase as acidified with ether (or chloroform, methylene dichloride) extraction, dehydrate organic phase, the powder of dry thing of gained and the dry gained of front organic phase merges.Use the lower alcohol dissolved powders, in solution, add lead acetate, remove the post precipitation concentrated filtrate, obtain the bilobalide crude product.After being dissolved in the crude product bilobalide in the organic solvent,,, obtain pure product bilobalide with carrying out the low temperature recrystallization after alcohol or the ketone dissolving by gac, silica gel adsorption column, concentrated effluent liquid.
Day disclosure special permission communique, flat 3-264533 (1991) has introduced solvent diacolation-extraction-crystallization-column chromatography.It is as follows that it extracts the bilobalide technical process:
---------------concentrated------centrifugal------------------------concentrated---------solution---cools off---crystallization---column chromatography separation---bilobalide to 50% ethanol to residue to gac to ethyl acetate/normal hexane to filtrate to diluent to extract to 60% acetone to Ginkgo Leaf in filtration in stirring to get organic phase in extraction in filtration in diafiltration.
In this technology, do not use the bigger chloroform of toxicity, tetracol phenixin equal solvent extracting process weeding of grease dissolubility detrimental impurity, but distillate extraction solvent (the residual solvent amount is no more than 10%) in the extracting solution earlier, after thin up and the cooling, the species precipitate that makes chlorophyll, derivative of fatty acid, biflavone etc. be insoluble in water is come out, and is removed by centrifugal method.
Also have some other solvent extraction or column chromatography separation method, principle is similar to be repeated no more.
ZL98111546.2 discloses a kind of " the crystalization in supercritical fluid separation method of material composition ", uses supercritical CO
2Can separate and obtain high-purity Ginkgolides.This method is initiatives such as applicant and has obtained Chinese invention patent.
Existing approximate density of liquid of supercutical fluid and dissolving power, the spread coefficient and the viscosity of the approximate gas of tool are called as " super solvent " again.The dissolving power of supercutical fluid produces significant difference with the difference of pressure, temperature and contained entrainment agent.The small size variation of pressure, temperature and entrainment agent can cause the strong variations of solute solubility.This invention is exactly this characteristic of utilizing supercutical fluid, crystallizer is set in extraction kettle, on the basis of choose reasonable entrainment agent, temperature and pressure by the control work system wait the solubleness that changes solute, allow crystallizable composition and noncrystalline component separating in the extract stream, realize the purifying of Ginkgolides, accompanying drawing 1 is seen in its technical process.
This method is compared with the conventional solvent extraction process, has simple to operately, and solvent consumption is little, environmental friendliness, production efficiency advantages of higher.
More above-mentioned each separation method, obviously:
In solvent extraction-lead acetate precipitation-silica gel adsorption-recrystallization method, because of using lead salt, easily cause heavy metals exceeding standard in the product, its aftertreatment complexity, what obtain only is Ginkgolides, and product purity is not high, inconvenience is adopted in the raw material pharmaceutical production.
Solvent diacolation-extraction-crystallization-column chromatography, its diacolation operation is unfavorable for suitability for industrialized production, and efficient is lower, and column chromatography is separated the back and is lacked the measure that is further purified, so stream part collection requirement is extremely tight, otherwise is difficult to obtain than straight product.
Supercritical CO
2Extractive crystallization process can be used for separating multiple product.For example, can realize Ginkgo Leaf → high-quality GBE according to the production purpose and the requirement of producer; GBE crude product → bilobalide, bilobalide and total lactone; The suitability for industrialized production of GBE crude product → qualified GBE product.But if produce ginkgolide monomer, still need supporting refining pure metallization processes and equipment with this method.
In existing each method, do not guarantee that all the monomeric crystal formation of gained is single, and its physicochemical property of the monomer of different crystal forms and effect also often there is difference about the bilobalide separation and purification.
Three, summary of the invention
The applicant has invented separation and purification bilobalide-like monomer methods in Ginkgolides, and Here it is uses column chromatography separation-recrystallization-supercritical CO
2Refining purifying and crystal formation arrangement combination process are to produce the bilobalide-like monomer of content 〉=90%.
The invention is characterized in Ginkgolides is loaded on column chromatography, the mixed solvent gradient elution is collected each monomer higher concentration stream part respectively, concentrates, and recrystallization is at last with the coarse-grain supercritical CO
2Make with extra care the arrangement of purifying and crystal formation.
Because the preparation technology of Folium Ginkgo extract (GBE) is ripe, and existing a large amount of GBE supplies on the market.With GBE (by version Chinese Pharmacopoeia specified standards in 2000) is initial feed, adopts supercritical fluid extraction crystallization process or solvent extration can make Ginkgolides.Technology of the present invention is starting raw material with the Ginkgolides, produces high-purity (content 〉=90%) Ginkgolide A, Ginkgolide B, ginkalide C, bilobalide J, bilobalide M and bilobalide.Its operational path is seen accompanying drawing 2.
Its stationary phase of the used column chromatography of the present invention includes but not limited to purification on normal-phase silica gel, reverse phase silica gel, aluminum oxide, metal-chelating resin, coordinate chromatograph filler etc.
Column chromatography is separated the employing gradient elution.The gradient elution solvent includes but not limited to water, ethanol, methyl alcohol, acetone, ethyl acetate, Virahol, acetic acid, sherwood oil etc.Each solvent change scope is from 1～99% in the mixed solvent.
Macrocrystalline supercritical CO
2The disclosed Crystallization Separation method of ZL98111546.2 of refining purifying and crystal formation arrangement request for utilization people invention.Supercritical CO
2Refining purifying and crystal formation finishing technique condition are pressure≤100MPa, temperature≤150 ℃, and entrainment agent is ethanol or ethyl acetate etc.
The advantage of this law is that technology is simple and direct, production is efficient, crystal formation is consistent, multiple high-purity ginkgolide monomer of no solvent residue.
For example: utilize the inventive method to produce Ginkgolide A, B, bilobalide, total yield 〉=90%, the high-content of monomer reaches 99.8%, can satisfy the specification of quality of chemical composition of Chinese materia medica standard reference material.
Four, description of drawings
Shown in Figure 1 is supercritical fluid extraction crystallization processes schema.
Shown in Figure 2 is present method process route chart.
Five, embodiment
Be example with separation and purification Ginkgolide A, Ginkgolide B and bilobalide in Ginkgolides now, non-limiting examples is described below:
Embodiment one:
Being added in the silica gel column chromatography after getting the Ginkgolides 100g drying of content 〉=97%, is moving phase (ratio of mixture is 90: 10～10: 90) with the petroleum ether-ethyl acetate mixed solvent, and gradient elution is collected Ginkgolide B (GB) stream part of purity 〉=50%; Vacuum concentration GB stream part gets the GB crude product to doing; With ethanol water recrystallization; Use supercritical CO again
2Refining purifying (32～100 ℃ of service temperatures, pressure 10～100MPa, extractive crystallization) is to remove trace impurity and solvent and to regulate and control crystal formation; Dry; Content greater than GB15～25g (with the content difference of GB in the Ginkgolides raw material, yield changes to some extent) of 99%; Total recovery 〉=90%.
Embodiment two:
Get the Ginkgolides 100g of content 〉=97%.Being added to after the drying in the silica gel column chromatography, is moving phase (ratio of mixture is 90: 10～10: 90) with the petroleum ether-ethyl acetate mixed solvent, gradient elution, Ginkgolide A (GA) the stream part of collecting purity 〉=50%.Vacuum concentration GA stream part gets the GA crude product to doing.With ethanol-acetic acid-water recrystallization, use supercritical CO again
2Refining purifying (32～150 ℃ of service temperatures, pressure 10～100MPa), removing trace impurity and solvent, and the regulation and control crystal formation; Dry; Can get high-content and be GA20～25g (with the content difference of GA in the Ginkgolides raw material, yield changes to some extent) of 99.8%; Total recovery 〉=85%.
Embodiment three:
Get the Ginkgolides 100g of content 〉=97%.Being added to after the drying on the reversed-phase silica gel column chromatography, is moving phase (ratio of mixture is 95: 5～5: 95) with the methanol-water mixed solvent, gradient elution, Ginkgolide B (GB) the stream part of collecting purity 〉=50%.Vacuum concentration GB stream part gets the GB crude product to doing.With first alcohol and water recrystallization, use supercritical CO again
2Refining purifying (32～150 ℃ of service temperatures, pressure 10～100MPa), removing trace impurity and solvent, and the regulation and control crystal formation; Dry; Content greater than GB25～35g (with the content difference of GB in the Ginkgolides raw material, yield changes to some extent) of 99%; Total recovery 〉=90%.
Embodiment four:
Get the Ginkgolides 100g of content 〉=97%.Being added to after the drying on the reversed-phase silica gel column chromatography, is moving phase (ratio of mixture is 95: 5～5: 95) with the methanol-water mixed solvent, gradient elution, Ginkgolide A (GA) the stream part of collecting purity 〉=50%.Vacuum concentration GA stream part gets the GA crude product to doing.With first alcohol and water recrystallization, use supercritical CO again
2Refining purifying (32～150 ℃ of service temperatures, pressure 10～100MPa), removing trace impurity and solvent, and the regulation and control crystal formation; Dry; Content greater than GA20～35g (with the content difference of GA in the Ginkgolides raw material, yield changes to some extent) of 99%; Total recovery 〉=90%.
Embodiment five:
Get the Ginkgolides 100g of content 〉=97%.Being added to after the drying on the reversed-phase silica gel column chromatography, is moving phase (ratio of mixture is 95: 5～5: 95) with the ethanol-water mixed solvent, gradient elution, bilobalide (BB) the stream part of collecting purity 〉=50%.Vacuum concentration BB stream part as for, the BB crude product.With first alcohol and water recrystallization, use supercritical CO again
2Refining purifying (32～150 ℃ of service temperatures, pressure 10～100MPa), removing trace impurity and solvent, and the regulation and control crystal formation; Dry; Content greater than BB 30～50g (with the content difference of BB in the Ginkgolides raw material, yield changes to some extent) of 97%, total recovery 〉=90%.
Claims (5)
1, the monomeric separation purification method of a kind of bilobalide-like is characterized in that: use column chromatography-recrystallization-supercritical CO
2Refining purifying combination process.Here it is at first separates Ginkgolides with column chromatography, the mixed solvent gradient elution is collected each monomer higher concentration stream part respectively, and concentrated then, recrystallization are at last with the coarse-grain supercritical CO
2Make with extra care the arrangement of purifying and crystal formation, control pressure≤100MPa, temperature≤150 ℃.
2, method according to claim 1 is characterized in that: the bilobalide-like monomer comprises Ginkgolide A, Ginkgolide B, ginkalide C, bilobalide J, bilobalide M and bilobalide.
3, method according to claim 1 is characterized in that: the column chromatography stationary phase comprises purification on normal-phase silica gel, reverse phase silica gel, aluminum oxide, metal-chelating resin, coordinate chromatograph filler.
4, according to claim 1 or 3 described methods, it is characterized in that: the gradient elution solvent comprises water, ethanol, methyl alcohol, acetone, ethyl acetate, Virahol, acetic acid, sherwood oil, and each solvent change scope is from 1～99% in the mixed solvent.
5, method according to claim 1 is characterized in that: adopt supercritical CO
2Carry out the arrangement of macrocrystalline refining purifying and crystal formation, pressure≤100MPa, temperature≤150 ℃, entrainment agent is ethanol, ethyl acetate etc., gained monomer content 〉=90% and crystal formation are single.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2004100651047A CN1634925B (en) | 2004-10-21 | 2004-10-21 | Separation and purification method for ginkgolide monomer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2004100651047A CN1634925B (en) | 2004-10-21 | 2004-10-21 | Separation and purification method for ginkgolide monomer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1634925A true CN1634925A (en) | 2005-07-06 |
| CN1634925B CN1634925B (en) | 2010-04-28 |
Family
ID=34846439
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2004100651047A Expired - Fee Related CN1634925B (en) | 2004-10-21 | 2004-10-21 | Separation and purification method for ginkgolide monomer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1634925B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100363023C (en) * | 2005-11-10 | 2008-01-23 | 济南大学 | Process for comprehensive treating root of kudzu vine actived element column chromatographic eluant |
| CN102911184A (en) * | 2012-11-14 | 2013-02-06 | 成都百裕科技制药有限公司 | Separation and purification preparation method for bilobalide |
| CN103044442A (en) * | 2013-01-03 | 2013-04-17 | 苏州泽达兴邦医药科技有限公司 | Method for separating and purifying GA, GB and bilobalide from folium ginkgo extractive |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1058628C (en) * | 1997-08-22 | 2000-11-22 | 孙云鹏 | Method for extracting flavone and terlactone from ginkgo leaf by supercritical carbon dioxide |
| CN1073867C (en) * | 1998-10-23 | 2001-10-31 | 潘见 | Separation method for material composition by supercritical fluid crystallization |
| US6117431A (en) * | 1999-12-03 | 2000-09-12 | Pharmline Inc. | Method for obtaining an extract from ginkgo biloba leaves |
-
2004
- 2004-10-21 CN CN2004100651047A patent/CN1634925B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100363023C (en) * | 2005-11-10 | 2008-01-23 | 济南大学 | Process for comprehensive treating root of kudzu vine actived element column chromatographic eluant |
| CN102911184A (en) * | 2012-11-14 | 2013-02-06 | 成都百裕科技制药有限公司 | Separation and purification preparation method for bilobalide |
| WO2014075461A1 (en) * | 2012-11-14 | 2014-05-22 | 成都百裕科技制药有限公司 | Separation and purification method for bilobalide |
| CN102911184B (en) * | 2012-11-14 | 2015-03-18 | 成都百裕科技制药有限公司 | Separation and purification preparation method for bilobalide |
| CN103044442A (en) * | 2013-01-03 | 2013-04-17 | 苏州泽达兴邦医药科技有限公司 | Method for separating and purifying GA, GB and bilobalide from folium ginkgo extractive |
| CN103044442B (en) * | 2013-01-03 | 2015-08-19 | 苏州泽达兴邦医药科技有限公司 | A kind of method of separation and purification GA, GB and bilobalide from Folium Ginkgo extract |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1634925B (en) | 2010-04-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11267775B2 (en) | Method for preparing cannabidiol by separation and purification using high-speed countercurrent chromatography | |
| CN101134758B (en) | Method for extracting and separating bilobalide A, B, C, J and bilobalide monomer from ginkgo leaf | |
| CN102976909B (en) | Method for extracting and purifying 6-gingerol from ginger | |
| CN100497344C (en) | Preparation of high-purity ginkgolide | |
| CN102351824B (en) | Method for preparing lactuca indica and lactucin | |
| CN104327127B (en) | Method for preparing angroside C, aucubin and harpagide through separation and purification by high-speed countercurrent chromatography | |
| CN104892687A (en) | Method for separating and purifying monomeric compound from Chinese mahonia leaves through high-speed counter-current chromatography | |
| CN104130226A (en) | Preparation method of high-content salvianolic acid B | |
| CN1171857C (en) | Process for enriching and purifying capsaicin with macroporous adsorption resin | |
| CN1394870A (en) | Method for separating and purifying tanshinone | |
| CN1634925B (en) | Separation and purification method for ginkgolide monomer | |
| CN104163754A (en) | Method for extraction and separating of high purity resveratrol from peanut root | |
| CN100500689C (en) | Method of separating and preparing wild jujube seed saponin | |
| CN102190665B (en) | Method for separating and purifying artemisinin by activated charcoal column chromatography employing nonaqueous system | |
| CN112159440B (en) | Phenolic glycoside compound and preparation method and application thereof | |
| CN1730027A (en) | Method for preparing anti-hepatitis active part from swertia main pharmaceutical plant | |
| CN109369382B (en) | A method for preparing ginkgolic acids | |
| CN102093210A (en) | Purified preparation method of six ginkgoic acid monomers | |
| CN103242390A (en) | Method for extracting methyldeactylasperulosidate and Scandoside methyl ester | |
| CN105061458B (en) | Preparation method of nontoxic high-purity ginkgo total lactones and monomers thereof | |
| CN113440547A (en) | Method for separating and purifying Japanese thistle herb total glycosides by adopting macroporous resin series dynamic axial compression column | |
| CN1194985C (en) | Diterpene compound of veronicastrum sibiricum and its extraction separation method | |
| CN1442413A (en) | Method of extracting medical taxadol and its derivative using regenerable resources | |
| CN1615995A (en) | Cyclodextrin super molecular composition of Chinese medicine giant knutweed rhizome and its preparing method | |
| CN1392130A (en) | Extracting and purifying method for hypericum perforatum component in plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100428 Termination date: 20161021 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |